LOCUS       sequence-406990-        6318 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhancer containing plasmid used for assessing enhancer::promoter 
            interactions.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6318)
  AUTHORS   DeMeis JD, Roberts JT, Delcher HA, Godang NL, Coley AB, Brown CL, 
            Shaw MH, Naaz S, Dahal A, Alqudah SY, Nguyen KN, Nguyen AD, Paudel 
            SS, Shell JE, Patil SS, Dang H, O'Neal WK, Knowles MR, Houserova D, 
            Gillespie MN, Borchert GM
  TITLE     Long G4-rich enhancers target promoters via a G4 DNA-based 
            mechanism.
  JOURNAL   Nucleic Acids Res. 2025 Jan 11;53(2):gkae1180. doi: 
            10.1093/nar/gkae1180.
  PUBMED    39658038
REFERENCE   2  (bases 1 to 6318)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2025-01-11"; volume: "53(2):gkae1180. doi"; 
            pages: " 10.1093/nar/gkae1180"
COMMENT     Sequence Label: TOPO-LG4_chr5:550kb antisense
FEATURES             Location/Qualifiers
     source          1..6318
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(143..148,149..166,167..173)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     186..208
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     205..221
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     205..221
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2751..2770)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2752..2770)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2777..2794)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2777..2793)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2786..2808)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2934..3362
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3021..3040)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3231..3252
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     CDS             3706..4500
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3760..3779)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4370..4389
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     CDS             join(4518..4586,4587..5378)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5549..6137
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6038..6057
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6291..6308
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagcttg
      241 gtaccgagct cggatccact agtaacggcc gccagtgtgc tggaattcgc ccttgggtgg
      301 atcacaaggt caggaatcaa gaccatctgg ttaacaccat gaaaccctgt ctcttaaaaa
      361 tatgaaaact tagccgggcg tggtggcagg cgcctgtagt cccagctact ggggggaggc
      421 cgaggcagga gaatggcgtg aacctgggag gcggagcttg cagtgagccg agatcacatc
      481 actgcactcc agcctgcgcc acagagcaaa actccgtcac acacacacaa aaaaatggta
      541 gctcctgagc caacactttt ccgctccccc acccgctgcg tctcagcatt ccttaggctc
      601 tccccttggg agactcacat gtatggggcg ggggagcggg ggtctcctag gccctgactc
      661 agctagagcg tctggctgac ccgtcattgt ccagggaagc catggatctg gtggccgcag
      721 cccagcgccg tcctcaccac ccaggcctgc ctggagccca gtggctgtca gggcagccga
      781 gaccagaagc gcaggtggag ccaaggcccc gctgcagtcc gcagagttgc aactctggga
      841 gctgagccct cacaagcctt gcttttcacc tcctcccagt gctaccagcc agaccccgag
      901 ggcccgggac cctccatgta cccatccaca cttctcggcc cacccctccc tgacagcccc
      961 ggccccccgg gagggagcag gtgcgtcccg cttcgggccc tctgaacccc ctccatttcg
     1021 ggctgtttca gacttcggcc tctctgaagc tcctccattt ctggctgctt ttcactgccg
     1081 tctgcctgcc tgtgttcttg tcttcacgcc ggcctttggt acgggaggga gccatccacg
     1141 gcccacactg cagctcccgt gacctccgca cggccccgtg cggacactca tctggatttt
     1201 gtttgtgctc aggagaggct ctgcattgaa cgaccttgca ctgggccggt ttcttttcta
     1261 aaggttccag agggcatggc tgtcaacttc ccgcttgtgt ccccgtcggg ttccagggca
     1321 cagaggttta tgcggagggc tcgagggact cacacctggg ggcagagttg ggcagcggcg
     1381 acgggcactc tggagcccat cacccccttg gcgtcgaggc ccaggggcct gatgtgccca
     1441 ctgctggcca gtttttgggt cgggcccctc ccccaggggg gtgctgggct accctcctga
     1501 gggcagaagc aattgttaag agcctcatca ctgtggtgtc agccactagc cctccccagc
     1561 tgtggaacag gggttggtcc ccatgcggag gggtcctggt cctgggcggc cccaccacac
     1621 cctgaaccgc ctgagcttct gactgcatgg aatattcaca ggactgcagt gtgagaaggc
     1681 agcaccgccc cggcgcccgc gaggaagccg aactgctcat aagttaagca gccccagcca
     1741 ggtcacgggg caaggaagcc gcctgcgacc tcggaaaccc ctggttcatc ccatgaccca
     1801 cccccgttca tccggtgacc cacccctcac cccggttcat cccgtgaccc acccccccgt
     1861 tcatcccatg acccagcccc ccaccccggt ttatcccgtg acccaccccc tgttcatccc
     1921 gtgacccacc cccgggtcat cctgtgaccc agaccccacc cctggtttat cccgtgaccc
     1981 accctcgggt catcccgtga cccagacccc acgcctggtt tatcccgtga cccacccgca
     2041 gcaccagcac ctcagtgctg cttttgtatt tatagtcgcc ttgctgtatg ttgagcgtaa
     2101 ttttacaatc tgcccaaaat tcctttgaag agcagatagg tgtgtacacc ctaaatgaat
     2161 aaaatacagt gcgatttggt gaaaggtaac ggctcgttta ttttccttgc cataaaaatg
     2221 ctacaatcac cctgaaggtg aacctggcaa gcggcatagg ggccagggcc tcactcggct
     2281 ctcctgtgca ccccgccccg gtgtgatgtt tggagatttg catacgaggc tctcagccag
     2341 gaaggaaatg gccaggaatc ccagaacgcg gcactgattt tccccatgtt atctgactcc
     2401 tgatattccc cgaactgtgt gaccactgcc tgtagttcag tctgtaatca aatactttgt
     2461 aaagactgta caagtgatcc cagacccaaa aggaaacgtc agaaatgcga ggctgtaggc
     2521 ttgttagaga tgggaggctg taggtttgtt agagatggga ggctgtaggc ttgttagaga
     2581 tgggaggctg ggctgtaggc ttgttagaga tgtgaggttg tagctgcagt tttcattttt
     2641 ctccagctga cgctgtcacc tgttgccatc agaaatctct caagggcgaa ttctgcagat
     2701 atccatcaca ctggcggccg ctcgagcatg catctagagg gcccaattcg ccctatagtg
     2761 agtcgtatta caattcactg gccgtcgttt tacaacgtcg tgactgggaa aaccctggcg
     2821 ttacccaact taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag
     2881 aggcccgcac cgatcgccct tcccaacagt tgcgcagcct gaatggcgaa tggacgcgcc
     2941 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     3001 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     3061 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     3121 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     3181 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     3241 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     3301 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3361 ttttaacaaa attcagggcg caagggctgc taaaggaagc ggaacacgta gaaagccagt
     3421 ccgcagaaac ggtgctgacc ccggatgaat gtcagctact gggctatctg gacaagggaa
     3481 aacgcaagcg caaagagaaa gcaggtagct tgcagtgggc ttacatggcg atagctagac
     3541 tgggcggttt tatggacagc aagcgaaccg gaattgccag ctggggcgcc ctctggtaag
     3601 gttgggaagc cctgcaaagt aaactggatg gctttcttgc cgccaaggat ctgatggcgc
     3661 aggggatcaa gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     3721 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     3781 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     3841 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     3901 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     3961 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatcc
     4021 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4081 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4141 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4201 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4261 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4321 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4381 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4441 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4501 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4561 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4621 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4681 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4741 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     4801 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     4861 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     4921 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     4981 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5041 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5101 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5161 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5221 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5281 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5341 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5401 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5461 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5521 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5581 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5641 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     5701 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     5761 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     5821 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     5881 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     5941 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6001 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6061 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6121 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6181 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     6241 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     6301 tgagcgagga agcggaag
//
